Vitamin D analogues

ABSTRACT

Vitamin D compounds formula I                    
     wherein R represents hydrogen, or R represents (C 1 -C 6 )alkyl, phenyl, or (C 7 -C 9 )aralkyl, optionally substituted with one or more groups selected from (C 1 -C 3 )alkyl, F, phenyl; n is an integer having the value 0, 1, or 2; and X represents hydroxy or halogen.

This application is the national phase under 35 U.S.C. § 371 of PCTInternational Application No. PCT/DK00/00389 which has an Internationalfiling date of Jul. 11, 2000, which designated the United States ofAmerica and was published in English and claims the benefit ofProvisional application No. 60/147,200, filed Aug. 4, 1999.

FIELD OF THE INVENTION

This invention relates to a hitherto unknown class of compounds thatshow strong activity in inducing differentiation and inhibitingundesirable proliferation of certain cells, including skin cells andcancer cells, as well as immunomodulating and antiinflammatory effects,to pharmaceutical preparations containing these compounds, to dosageunits of such preparations, and to their use in the treatment and/orprophylaxis of diseases characterized by abnormal cell differentiationand/or cell proliferation.

BACKGROUND OF THE INVENTION

A number of vitamin D analogues have been described that show somedegree of selectivity in favour of the cell differentiationinducing/cell proliferation inhibiting activity in vitro as compared tothe effects on calcium metabolism in vivo (as measured in increasedserum calcium concentration and/or increased urinary calcium excretion),which adversely limit the dosage that can safely be administered topatients. One of the first of these to appear, calcipotril (INN) orcalcipotriene (USAN), has been developed on the basis of thisselectivity and is now recognized worldwide as an effective and safedrug for the topical treatment of psoriasis.

A study with another vitamin D analogue, seocalcitol, selected on thisbasis supports the concept that systemically administered vitamin Danalogues may inhibit breast cancer cell proliferation in vivo atsub-toxic doses (Colston, K. W. et al., Biochem. Pharmacol. 44,2273-2280 (1992)).

Another vitamin D analogue, CB1093 (20-epi-22-ethoxy-23-yne-24a, 26a,27a, trishomo-1α,25(OH)₂D₃ vitamin D₃) (Calverley M. J. et al. In:Vitamin D, Proceedings of the Ninth Workshop on Vitamin D, Orlando,Fla., Walter de Gruyter, Berlin, 1994, pp 85-86; and disclosed in WO93/19044) has been shown to possess potent activity in an in vitro assayon inhibiting the invasiveness of human carcinoma cells (Hansen C. M. etal. In: Vitamin D, Proceedings of the Ninth Workshop on Vitamin D,Orlando, Fla., Walter de Gruyter, Berlin, 1994, pp 508-509).

CB1093 has also been demonstrated to have potent inhibitory activity onthe proliferation of, and stimulatory activity on the differentiationand apoptosis of, different types of cancer cells, such as, brain glialtumor cells in vitro (Baudet, C. et al., Cancer Lett. 1996, 100, 3);MCF-7 breast cancer cells in vitro and in vivo (Colston, K. W., et al.,In: Vitamin D, Proceedings of the Tenth Workship on Vitamin D,Strasbourg, France, 1997, University of California, Riverside, 1997, pp443-450; Danielsson, C. et al., In: Vitamin D, Proceedings of the TenthWorkshop on Vitamin D, Strasbourg, France, 1997, University ofCalifornia, Riverside, 1997, pp 485-486; Danielsson, C. et al., J.Cellular Biochem., 1997, 66, 552); NB4 acute promyelocytic leukemiacells in vitro (Elstner, E., et al., J. Clin. Invest., 1997, 99, 349);HL-60 and de novo human acute myeloid leukemia cells in vitro (Pakkala,I. et al., Blood 1995, 86(10, Suppl.), 775a; Pakkala, I. et al.,Leukemia Research 1997, 21, 321); and MG-63 human osteosarcoma cells invitro (Ryhänen, S., et al., J. Cellular Biochem. 1998, 70, 414).

CB 1093 also significantly decreased plasma PTH and phosphate levels inchronically uraemic rats with secondary hyperparathyroidism (Hruby, M.et al., Nephrol. Dial. Transplant. 1996, 11, 1781).

The classical calcemic vitamin D activity of CB1093, as determined bythe urinary excretion of calcium in rats, has been determined to 27% ofthat of 1α,25(OH)₂D₃ and the calcemic activity of seocalcitol in thesame assay has been determined to 50% (Danielsson, C. et al., J.Cellular Biochem., 1997, 66, 552). In an in vivo experiment treatingrats with mammary tumours with CB1093 (1 μg/kg body weight for 28 days )there was a 49% reduction of the initial tumour volume, but there wasstill a slight increase in the serum calcium concentration (ibid.). Thisindicates that the therapeutic window may still be rather narrow, andconcern about possible induced increases in serum calcium levels cannotyet be excluded.

Another problem using vitamin D analogues in the non-topical treatmentof hyperproliferative diseases, such as cancer, is metabolic stabilityin vivo. This stability has to be above a certain minimum level, for acompound to be used in practical therapy. As shown in table 1, thestability of CB1093 in an in vitro-model of metabolic stability, usingrat liver homogenate “S-9” (Kissmeyer, A.-M. et al., Biochem.Pharmacol., 1997, 53, 1087) is quite low compared to seocalcitol (T½ 1.3hr) and 1α,25(OH)₂D₃ (T½ 2.5 hr).

There is therefore a continuing need for new vitamin D analogues withhigh anti-cell proliferative and/or cell differentiation inducingactivity showing an acceptable combination of prolonged therapeuticactivity and minimum toxic effects compared to 1α,25(OH)₂D₃. The purposeof the present invention is to provide such new compounds, which purposeis achieved with the novel compounsd having the general formula Iherein.

SUMMARY OF THE INVENTION

The compounds of the invention constitute a novel class of vitamin Danalogues represented by the general formula I:

wherein formula R represents hydrogen, or (C₁-C₆)alkyl, phenyl, or(C₇-C₉)aralkyl optionally substituted with one or more of (C₁-C₃)alkyl,F, or phenyl; n is an integer having the value 0, 1, or 2; and Xrepresents hydroxy or halogen.

DETAILED DESCRIPTION OF THE INVENTION

Preferred embodiments of the invention.

Preferred compounds of formula I are compounds wherein R representsmethyl, ethyl, propyl, isopropyl, benzyl, and ortho methylbenzyl, metamethylbenzyl, and para methylbenzyl. More preferably R represents methylor ethyl. n is preferably 0 or 1; 1 being more preferred. Preferably Xrepresents OH, F, or Cl. Most preferably X represents F or X mostpreferably represents OH or Cl.

The compounds of the invention can comprise more than onediastereoisomeric form; that is both R and S configurations at thecarbon atoms marked 22, 25 and 26 in formula I. The invention covers allthese diastereoisomers in pure form and also mixtures thereof. Preferredisomers are compounds having the configurations 22(S),25(S),26(S) and22(S),25(S),26(R). In addition, prodrugs of compounds of formula I inwhich one or more of the hydroxy groups are masked as groups that can bereconverted to hydroxy groups in vivo could also be envisaged.

The compounds I may be obtained in crystalline form either directly byconcentration from an organic solvent or by crystallisation orrecrystallisation from an organic solvent or mixture of said solvent anda co-solvent which may be organic or inorganic, such as water. Thecrystals may be isolated in essentially solvent-free form or as asolvate, such as a hydrate. The invention covers all crystallinemodifications and forms and also mixtures thereof.

Exemplary compounds of the invention are

1(S),3(R)-Dihydroxy-20(R)-(5-ethyl-1(S),5(S),6(S)-trihydroxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 101),

1(S),3(R)-Dihydroxy-20(R)-(5(S),6(S)-dihydroxy-5-ethyl-1(S)-methoxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 102),

1(S),3(R)-Dihydroxy-20(R)-(5(S),6(S)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 103),

1(S),3(R)-Dihydroxy-20(R)-(5(S),6(S)-dihydroxy-5-ethyl-1(S)-(1-propyloxy)-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 104),

1(S),3(R)-Dihydroxy-20(R)-(1(S)-benzylyloxy-5(S),6(S)-dihydroxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 105),

1(S),3(R)-Dihydroxy-20(R)-(5(R),6(S)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 106),

1(S),3(R)-Dihydroxy-20(R)-(5(R),6(R)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 107),

1(S),3(R)-Dihydroxy-20(R)-(5(S),6(R)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 108),

(S),3(R)-Dihydroxy-20(R)-(4-ethyl-1(S),4(S),5(S)-trihydroxy-2-hexyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 109),

(S),3(R)-Dihydroxy-20(R)-(4(S),5(S)-dihydroxy-1(S)-ethoxy 4 ethyl 2hexyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 111),

1(S),3(R)-Dihydroxy-20(R)-(4-ethyl-1(S),4(R),5(S)-trihydroxy-2-hexyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 114),

1(S),3(R)-Dihydroxy-20(R)-(4(R),5(S)-dihydroxy-1(S)-ethoxy-4-ethyl-2-hexyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(Compound 116),

1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy-5-ethyl-6(S)-fluoro-5(S)-hydroxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(compound 149), and

1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy-5-ethyl-6(R)-fluoro-5(S)-hydroxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(compound 150).

1(S),3(R)-Dihydroxy-20(R)-(1(S),5(R/S)-dihydroxy-5-ethyl-6(S)-fluoro-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(compound 157)

1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy5-ethyl-6(S)-fluoro-2-heptyn-5(R/S)-hydroxy1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 158)

1(S),3(R)-Dihydroxy-20(R)-(1(S),5(R/S)-dihydroxy-5-ethyl-6(R)-fluoro-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene(compound 159)

1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy5-ethyl-6(R)-fluoro-2-heptyn-5(R/S)-hydroxy1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 160)

As used in the specification the following terms have the meaningindicated:

“Alkyl” refers to any univalent group derived from an alkane by removalof a hydrogen atom from any carbon atom, and includes the subclasses ofnormal alkyl (n-alkyl), and primary, secondary and tertiary alkyl groupsrespectively, and having the number of carbon atoms specified, includingfor example (C₁-C₃)alkyl, (C₁-C₆)alkyl, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, and n-hexyl.Alkane refers to an acyclic straight or branched hydrocarbon having thegeneral formula C_(n)H_(2n+2,) and therefore consisting entirely ofhydrogen atoms and saturated carbon atoms.

“(C₇-C₉)aralkyl” refers to any alkyl group substituted with an aromaticgroup, such as phenyl, and having the total number of carbon atomsspecified, preferred are (C₇-C₈)aralkyl groups. Examples are benzyl,2-phenyl-ethyl, ortho methylbenzyl, meta methylbenzyl, and paramethylbenzyl.

“Halogen” means the same or different of fluoro, chloro, bromo, andiodo.

The present invention provides a hitherto undisclosed series of vitaminD analogues which is characterised by the presence of an additionalhydroxy group or halogen atom in the 26 position of the side chain.Compared to the prior art vitamin D analogues, illustrated by CB1093,the present new vitamin D analogues have precisely the properties whichare demanded (Table 1): Half or less the calcemic activity (in the ratcalciuric model) and much higher metabolic stability (in the S-9 ratliver homogenate model), together with an only slightly reducedantiproliferative activity (in two different cancer assays: The U937leukemia cell assay (Kissmeyer, A.-M. et al., Biochem. Pharmacol., 1997,53, 1087) and the MCF-7 mammary cancer cell assay (Danielsson, C. etal., J. Cellular Biochem., 1997, 66, 552)). Moreover the activity of thepresent Compounds I in the HaCaT assay, a psoriasis model (Kissmeyer,A.-M. et al., Biochem. Pharmacol., 1997, 53, 1087), is slightly higherthan that of CB1093. The advantageous properties of the 26-hydroxy or26-halogen vitamin D analogues of the present invention are entirelyunexpected, as it is known that a similar introduction of a 26-hydroxygroup in seocalcitol results in compounds with drastically reducedactivities in both the U937 and the HaCaT assays. All the four possible26-hydroxy-seocalcitol analogues have been shown to be naturalmetabolites of seocalcitol in vitro and in vivo in rats and in vitro inhumans (Binderup, E., et al., In: Vitamin D, Proceedings of the TenthWorkshop on Vitamin D, Strasbourg, France, 1997, University ofCalifornia, Riverside, 1997, pp 89-90; Kissmeyer, A.-M. et al., Biochem.Pharmacol., 1997, 53, 1087).

TABLE 1 Compound 25/26-Configur. No. 103 § No. 108 § CB1093 §, $ Assay1α, 25(OH)₂D₃ 25(S), 26(S) 25(S), 26(R) (No 26-OH) U937, -log IC₅₀ 7.5 ±0.3 9.0 ± 0.2 8.9 ± 0.2 9.5 ± 0.1 U937, rel.* 1.0 36 49 74 MCF-7, -logIC₅₀ 7.7 ± 0.3 9.9 ± 0.1 9.8 ± 0.2 10.2 ± 0.3  MCF-7, rel.* 1.0 153  89311 HaCaT, -log IC₅₀ 7.4 ± 0.3 9.0 ± 0.3 8.8 ± 0.2 8.7 ± 0.9 HaCaT,rel.* 1.0 38 30 18 S-9, metab. in vitr. #    0.43    0.29    0.02Calcem.; in vivo* 1.0    0.11    0.6    0.27 Notes to Table 1: § R =Ethyl, n = 1 and 22(S)-configuration $ Reference compound: No26-OH-group; otherwise the same structure as Compound 103/108 *Geometricmean of the ratios, relative to 1α, 25(OH)₂D₃, from all experiments withthe compound in question, in the assay concerned; the higher the value,the more potent the compound # Fraction left after 1 hour incubation

The following standard abbreviations are used throughout thisdisclosure:

AcOH=acetic acid

18C6=18-Crown-6

b.p.=boiling point

Bn=benzyl

Bu=n-butyl

Comp=Compound No.

DMAP=4-dimethylaminopyridine

DMF=N,N-dimethylformamide

Et=ethyl

EtOAc=ethyl acetate

Exam=Example No.

eqv=equivalent (molar)

Ether=diethyl ether

G.P.=General Procedure No.

Hal=Cl, Br or I

h=hour

Me=methyl

m.p.=melting point

Ms=methanesulfonate

PG=Protective Group

Ph=phenyl

Pr=n-propyl

Prep=Preparation No.

PPTS=pyridinium p-toluenesulfonate

Py=pyridine

TBAF=tetra-n-butylammonium fluoride

TBS=tert butyldimethylsilyl

Tf=trifluromethanesulfonyl

THF=tetrahydrofuran

THP=tetrahydro-4H-pyran-2-yl

TMS=trimethylsilyl

Tol=toluene

Ts=4-toluenesulfonyl

Compounds of formula I, as illustrated in Table 4, may be prepared bythe general method of Scheme 1:

Notes to Scheme 1:

General: X¹═O−PG², F, Cl, Br, or I

PG¹ and PG²=Hydrogen or Protective Groups: the same or different, or acombined bifunctional group.

Q=H or Me₃Si; n=1,2 or 3.

(a) CH₃NH—OCH₃, HCl; EtMgBr (3 eqv)

(b) i) Q-C≡C—(CH₂)_(n)-Met; (Met=−Li, −MgHal; −AlBr₂, −Br+SmI₂);

if it is desired (only for Q=H) that: PG¹=TMS and X¹═O-PG¹=O-TBS andQ=H;

ii) Me₃SiCl/Et₃N/CH₂Cl₂/DMAP,

or, if it is desired that: PG¹-PG²=—C(CH₃)₂—, i.e. X¹═O-PG²(-PG¹-), andQ=H:

ii) TBAF/THF, (→PG¹=PG²=Q=H)

iii) CH₂═C(CH₃)—O—CH₃ or (CH₃)₂C(OCH₃)₂/TsOH

or, if it is desired that: PG¹-TMS and X¹═F:

Compounds 509 or 510 (e.g. prepared as in Scheme 1a) replace compounds503 and 504 of Scheme 1;

or, if it is desired that: PG¹=TMS and X¹═F, Cl, Br, or I and Q=H:

ii) TBAF/THF, (→PG¹=PG²=Q=H, i.e. X¹═OH)

iii) Conversion of X¹ (═OH) to X¹═F, Cl, Br, or I, for example as shownin Scheme 2;

iv) optional conversion of PG¹=H to PG¹=H to PG¹=TMS, e.g. withMe₃SiCl/Et₃N/CH₂Cl₂/DMAP

(c) i) II+BuLi; ii) 1; iii) optional alkylation of 22-OH with RZ/base(Z=Good leaving group, e.g. Hal, Ms, Ts, or Tf)

(d) Triplet sensitized photo-isomerization of the vitamin D triene,5(6)(E) to 5(6)(Z); ii) optional alkylation of 22-OH with RZ/base

(e) i) Deprotection with HF/CH₃CN/EtOAc or TBAF/THF, optionally followedby or preceded by PPTS/EtOH.

Notes to Scheme 1a:

(a) KF, HCONH₂, 70° C. (method of Fritz-Langhals, E. et al, Tetr. Lett.1993, 34, 293)

(b) CH₃NH—OCH₃, HCl, EtMgBr (3 eqv)

Compounds I can be prepared from the vitamin D-derived aldehyde compound1, a synthesis of which has been reported (Calverley, M. J.,Tetrahedron, 1987, 43, 4609.), for example by the routes outlined inScheme 1, by reaction with an organometallic derivative of the sidechain building blocks of general formula II.

The Compounds II to be used for making the Compounds I where X═OH can besynthesized as follows:

L(−)Ethyl lactate or D(+)ethyl lactate (or the corresponding methylester) is protected by silylation with tert-butyldimethylsilyl chlorideto give the corresponding ethyl TBS-lactate (501, 502). This isconverted, in a direct procedure with ethyl magnesium bromide, to thecorresponding ethylketone, (503, 504), via the intermediaryN-methoxy-N-methylamides, by the method of Williams, J. M. et al. (Tetr.Lett., 1995, 36, 5461).

The ketone 503 or 504 is converted to the partially protected side chainsynthon of general formula II by reaction with an organometallic reagentof the type Q-C≡C—(CH₂)_(n)-Met; (Met=−Li, −MgHal; −AlBr_(2;) −Hal+e.g.Smi₂; Q=H or Me₃Di; n=1, 2 or 3). In the case ofQ-C≡C—(CH₂)_(n)-Hal+e.g. Smi₂, a Barbier type reaction between a halide,a ketone, and a metal/metal salt is referred to.

A mixture of two diasteroisomers is usually formed: The R and the Sisomer at the carbon atom marked (25) in Scheme 1, that is the carbonatom which ends up being the carbon atom C(25) in the final compound offormula I. If desired, the two diastereoisomers may be separated at thisstage, or later during the synthesis of the side chain synthons II, ifthis is more convenient.

The following steps in the synthesis of the fully protected side chainsynthons II are: 1) If Q is trimethylsilyl, Q is converted into hydrogenby deprotection, e.g. by means of a base. 2) The unprotected hydroxygroup at the carbon marked (25) is protected, e.g. by silylation withTMS-Cl; such that PG¹=TMS (and PG²=TBS).

Alternatively the TBS group (PG²) of compound II may be removed, e.g. byTBAF (if Q=TMS, this is converted to H at the same time). The twohydroxy groups at the carbon atoms marked (25) and (26), respectively,can then be protected in one step, by conversion into a cyclic acetal orketal, e.g. an acetonide (isopropylidene ketal), e.g. by means of e.g.2-methoxypropene or 2,2-dimethoxypropane and an acid, such that PG¹ andPG² are connected into one group: —C(CH₃)₂—. Other methods of protectionof 1,2-diols are described in the literature, e.g. in: Greene, T. W. andWuts, P. G. M., “Protective Groups in Organic Synthesis”, Sec. Ed., JohnWiley and Sons, New York 1991, pp 118-142. An advantage of using cyclicacetals or ketals, such as acetonides, as protective groups of compoundsII is that they are particularly well suited for establishing thestereochemistry at carbons (25) and (26) by way of Nuclear OverhauserEnhancement (NOE) NMR spectroscopy.

The Compounds II to be used for making the Compounds I where X═F may besynthesized from ketone 509 or 510, using the method of Scheme 1a andthe intermediates of Table 1a:

TABLE 1a Fluorinated Side Chain Building Blocks of Scheme 1a Prep CompG.P. C(26) 42 505 11 R 43 506 11 S 44 507 12 S 45 508 12 R 46 509 1a S47 510 1a R

The compounds II to be used for making the Compounds I where X is Cl,Br, or I are preferably made (and for X═F may be made) from thecorresponding Compounds II where O-PG¹ and O-PG² (=X¹) are both OH, forexample as shown in Scheme 2:

Note to Scheme 2

The compounds II where X¹═Cl or I can be converted to each other orconverted to the corresponding compounds where X¹═F or Br by standardmethods, e.g. as described in: R. C. Larock, Comprehensive OrganicTransformations, VCH Publishers, Inc., New York, N.Y., USA, 1989, pp.337-339 hereby incorporated by reference.

TABLE 2 Side Chain Building Blocks of General Formula II (Scheme 1) PrepComp G.P. Q n C(25) C(26) PG¹ PG² 4 201 2 H 1 S S H TBS 5 202 4 H 1 S SH H 6 203 5 H 1 S S —C(CH₃)₂— 7 204 6 H 1 S S TMS TBS 8 205 2 H 1 R S HTBS 9 206 4 H 1 R S H H 10 207 5 H 1 R S —C(CH₃)₂— 11 208 6 H 1 R S TMSTBS 12 209 2 H 1 R R H TBS 13 210 6 H 1 R R TMS TBS 14 211 2 H 1 S R HTBS 15 212 6 H 1 S R TMS TBS 16 213 3 H 0 S S H TBS 17 214 3a TMS 0 S SH TBS 18 215 4 H 0 S S H H 19 216 5a H 0 S S —C(CH₃)₂— 16 217 3 H 0 R SH TBS 17 218 3a TMS 0 R S H TBS 18 219 4 H 0 R S H H 20 220 5a H 0 R S—C(CH₃)₂— Note to Table 2: X¹ = O-PG²

TABLE 2a Fluorinated Side Chain Building Blocks of General Formula II(Scheme 1) Prep Comp G.P. Q n C(25) C(26) PG¹ X¹ 48 221 2a H 1 R/S S H F49 222 6 H 1 R/S S TMS F 50 223 2a H 1 R/S R H F 51 224 6 H 1 R/S R TMSF

The reaction of the aldehyde 1 with the organometallic reagents derivedfrom the side chain building blocks II, can be performed by standardmethods of nucleophilic addition of organometallic reagents to carbonylcompounds; i.e. by reacting the alkyne intermediate II with a Grignardreagent, such as ethyl magnesium bromide, or an alkyl lithium, such asbutyl lithium (General Procedure 7) in a suitable anhydrous solvent,such as ether and/or THF, to generate the metal acetylide, then adding1, to give III after usual aqueous work-up (which is normally implied inall the reactions of Scheme 1. In general the reaction product III is amixture of the two possible C-22 epimers, here designated IIIA and IIIB.It is usually preferable to separate the IIIA and IIIB epimers which canconveniently be done by chromatography.

Nonlimiting illustrations of such compounds of formula III are given inTables 3 and 3a. Compounds IIIA are formed in much higher yields thanthe corresponding IIIB epimers, typically in the ratio of about 95 to 5.Compounds IIIA, IVA and IA have 22(S) stereochemistry, and thecorresponding compounds with the suffix B have 22(R) stereochemistry.Compounds IA are the preferred ones.

The optional alkylation of the 22-hydroxy compounds of general formulaIII or IV to yield the corresponding compound III or IV where R is(C₁-C₆)alkyl, phenyl, or (C₇-C₉)aralkyl can be performed by standardmethods well known to the specialist. Illustrative, but non limiting,compounds of this sort are listed in Table 3.

In the alkylation reaction use is preferably made of an alkylating agentRZ, in which Z stands for a good leaving group, such as for example Hal,Ms, Tf; the RZ being allowed to react with the anion of the appropriatecompound III or IV (R═H), derived therefrom by means of a suitablestrong base, such as an alkali-metal alkoxide, alkyl alkali-metal oralkali-metal hydride. A suitable crown ether may be added as a phasetransfer agent to accelerate the alkylation process. A useful method isdescribed in General Procedure 9.

The photo-isomerization of the vitamin D triene, 5(6)(E), Compounds IIIof Scheme 1, to 5(6)(Z), Compounds IV of Scheme 1, is performed by meansof UV-light in the presence of a triplet sensitizer, e.g. anthracene;useful methods are described in General Procedure 8 and and 8a.Nonlimiting illustrations of such compounds of general formula IV aregiven in Tables 3 and 3a, along with references to the preparation ofeach compound.

The triplet sensitized photo-isomerization of the vitamin D triene,5(6)(E), Compounds III, to 5(6)(Z), Compounds IV, and the (optional)alkylation of the 22-OH-group with RHal/base, to form a 22-O—R compoundwhere R≠H, may be performed in arbitrary order, according to what ismost convenient in each case.

The final step in the synthesis of Compounds I of the present invention,examples of which are listed in Table 4, is one or more deprotectionprocedures to remove all protective groups of the compounds of generalformula IV of Scheme 1. The deprotection may for example be performedeither with TBAF to remove silyl groups, like TMS or TBS groups (GeneralProcedure 4), or with HF which removes both silyl groups and acidsensitive protective groups, such as isopropylidene (ketal) groups(General Procedure 10). Alternatively, if both types of protectivegroups are to be removed, two different selective procedures may be usedin sequence, e.g. as described in WO97/46522 (G.P. 7): The silyl groupsare removed with TBAF, followed by the use of PPTS which selectivelyremoves acid-sensitive protective groups; (or in the reverse order).

TABLE 3 Intermediates of General Formulas III and IV (Scheme 1) TypePrep Comp G.P. R n C(25) C(26) PG¹ PG² IIIA 21 301 7 H 1 S S TMS TBSIIIB 21 302 7 H 1 S S TMS TBS IVA 22 401 8 H 1 S S TMS TBS IVA 23 402 9Me 1 S S TMS TBS IVA 24 403 9 Et 1 S S TMS TBS IVA 25 404 9 Pr 1 S S TMSTBS IVA 26 405 9 Bn 1 S S TMS TBS IIIA 27 303 7 H 1 R S TMS TBS IVA 28406 8 H 1 R S TMS TBS IVA 29 407 9 Et 1 R S TMS TBS IIIA 30 304 7 H 1 RR TMS TBS IVA 31 408 8 H 1 R R TMS TBS IVA 32 409 9 Et 1 R R TMS TBSIIIA 33 305 7 H 1 S R TMS TBS IVA 34 410 8 H 1 S R TMS TBS IVA 35 411 9Et 1 S R TMS TBS IIIA 36 306 7a H 0 S S —C(CH₃)₂— IVA 37 412 8 H 0 S S—C(CH₃)₂— IVA 38 413 9 Et 0 S S —C(CH₃)₂— IIIA 39 307 7a H 0 R S—C(CH₃)₂— IVA 40 414 8 H 0 R S —C(CH₃)₂— IVA 41 415 9 Et 0 R S —C(CH₃)₂—Note to Table 2: X¹ = O-PG²

TABLE 3a Fluorinated Intermediates of General Formulas III and IV(Scheme 1) Type Prep Comp G.P. R n C(25) C(26) PG¹ X¹ IIIA 52 308 7 H 1R/S S TMS F IVA 53 416 8a H 1 R/S S TMS F IVA 54 417 9 Et 1 R/S S H FIIIA 55 309 7 H 1 R/S R TMS F IVA 56 418 8a H 1 R/S R TMS F IVA 57 419 9Et 1 R/S R H F

Exemplified Compounds I of the invention are listed in Table 4, thenumbered examples giving reference to illustrative methods of synthesis,together with spectroscopic data for the exemplified compounds.

The Compounds, 110, 112-113, 115 and 117-156 are made in a sequence ofsynthetic steps which is analogous to the sequence used for thepreparations of Compounds, 101-109, 111, 114, 116 and 157-160, Examples1-12 and 16-19:

Compound 1 and the appropriate side chain building blocks of GeneralFormula II, H—C≡C—(CH₂)_(n)—C(C₂H₅)(OPG¹)-CH(X¹)CH₃, are reacted,according to General Procedure 7 (G.P. 7), to give the correspondingcompound of formula III.

If not mentioned in Tables 2 or 2a, the compounds II in question can beprepared by similar methods to those applied for the synthesis of thecompounds II listed in Tables 2 or 2a.

The compound of formula III is photoisomerized, according to G.P. 8 or8a, to give the corresponding compound of formula IV.

Optionally, the compound of formula IV and the appropriate alkylatingagent RZ are reacted, according to G.P. 9, to give the correspondingcompound of formula IV where R≠H. The photoisomerization step andalkylation step may be performed in the reverse order, if desired.

As the last step, the compound of formula IV is deprotected, accordingto either G.P. 4 or G.P. 10 to give the Compound I in question.

TABLE 4 Compounds of General Formula I Type Exam Comp G.P. R n X C(25)C(26) IA 1 101 4 H 1 OH S S IA 2 102 4 Me 1 CH S S IA 3 103 4 Et 1 OH SS IA 4 104 4 Pr 1 OH S S IA 5 105 4 Bn 1 OH S S IA 6 106 4 Et 1 OH R SIA 7 107 4 Et 1 OH R R IA 8 108 4 Et 1 OH S R IA 9 109 10 H 0 OH S S IA110 10 Me 0 OH S S IA 10 111 10 Et 0 OH S S IA 112 10 Pr 0 OH S S IA 11310 Bn 0 OH S S IA 11 114 10 H 0 OH R S IA 115 10 Me 0 OH R S IA 12 11610 Et 0 OH R S IA 117 10 Pr 0 OH R S IA 118 10 H 0 OH R R IA 119 10 Et 0OH R R IA 120 10 H 0 OH S R IA 121 10 Me 0 OH S R IA 122 10 Et 0 OH S RIA 123 10 Pr 0 OH S R IA 124 10 H 1 OH R S IA 125 10 Me 1 OH R S IA 12410 H 1 OH R R IA 125 10 Me 1 OH R R IA 126 10 H 1 OH S R IA 127 10 Me 1OH S R IA 128 10 Pr 1 OH S R IA 129 10 Bn 1 OH S R 1A 131 10 H 2 OH S SIA 132 10 Me 2 OH S S IA 133 10 Et 2 OH S S IA 134 10 Pr 2 OH S S IA 13510 Bn 2 OH S S IA 136 10 Et 2 OH R S IA 137 10 Et 2 OH R R IA 138 10 H 2OH S R IA 139 10 Me 2 OH S R IA 140 10 Et 2 OH S R IA 141 10 Pr 2 OH S RIB 142 10 Et 1 OH S S IA 143 10 Me 0 F S S IA 144 10 Me 0 F S R IA 14510 Et 0 F S S IA 146 10 Et 0 F S R IA 147 10 Me 1 F S S IA 148 10 Me 1 FS R IA 149 10 Et 1 F S S IA 150 10 Et 1 F S R IA 151 10 Et 2 F S S IA152 10 Me 0 Cl S S IA 153 10 Et 0 Cl S S IA 154 10 Me 1 Cl S S IA 155 10Et 1 Cl S S IA 156 10 Et 1 Cl S R IA 16 157 10 H 1 F R/S S IA 17 158 10Et 1 F R/S S IA 18 159 10 H 1 F R/S R IA 19 160 10 Et 1 F R/S R

The present compounds are intended for use in pharmaceuticalcompositions which are useful in the local or systemic treatment orprophylaxis of human and veterinary disorders, such as e.g. psoriasis(including pustulosis palmoplantaris, acrodermatitis continua and nailpsoriasis) and other disturbances of keratinization, HIV-associateddermatoses, wound healing, various cancer forms, such as leukemia,mammary cancer, brain glial tumours, osteosarcoma, myelofibrosis,melanoma, other skin cancers, and of diseases of, or imbalances in, theimmune system, such as host versus graft and graft versus host reactionand transplant rejection, and autoimmune diseases, such as discoid andsystemic lupus erythematosus, diabetes mellitus and chronic dermatosesof auto-immune type, e.g. scleroderma and pemphigus vulgaris, andinflammatory diseases, such as asthma and rheumatoid arthritis, as wellas a number of other diseases states including hyperparathyroidism,particularly secondary hyperparathyroidism associated with renalfailure, cognituve impairment or senile dementia (Alzheimers disease)and other neurodegenerative diseases, hypertension, acne, alopecia, skinatrophy, e.g. steroid induced skin atrophy, skin ageing, includingphoto-ageing, and to their use for promoting osteogenesis andtreating/preventing osteoporosis and osteomalacia.

The present compounds may be used in combination with otherpharmaceuticals or treatment modalities. In the treatment of psoriasisthe present compounds may be used in combination with otherantipsoriatic drugs, e.g steroids, or with other treatments e.g. light-or UV-light-treatment or the combined PUVA-treatment. In the treatmentof cancer the present compounds may be used in combination with otheranti-cancer drugs or anti-cancer treatments, such as radiationtreatment. In the prevention of graft rejection and graft versus hostreaction, or in the treatment of auto-immune diseases, the presentcompounds may advantageously be used in combination with otherimmunosuppressive/immunoregulating drugs or treatments, e.g. withcyclosporin A.

The amount required of a compound of formula I (hereinafter referred toas the active ingredient) for therapeutic effect will, of course, varyboth with the particular compound, the route of administration and themammal under treatment. The compounds of the invention can beadministered by the parenteral, intra-articular, enteral or topicalroutes. They are well absorbed when given enterally and this is thepreferred route of administration in the treatment of systemicdisorders. In the treatment of dermatological disorders like psoriasisor eye diseases topical or enteral forms are preferred.

While it is possible for an active ingredient to be administered aloneas the raw chemical, it is preferable to present it as a pharmaceuticalformulation. Conveniently, the active ingredient comprises from 0.1 ppmto 0.1% by weight of the formulation.

The formulations, both for veterinary and for human medical use, of thepresent invention thus comprise an active ingredient in association witha pharmaceutically acceptable carrier therefore and optionally othertherapeutic ingredient(s). The carrier(s) must be “acceptable” in thesense of being compatible with the other ingredients of the formulationsand not deleterious to the recipient thereof.

The formulations include e.g. those in a form suitable for oral,ophthalmic, rectal, parenteral (including subcutaneous, intramuscularand intravenous), transdermal, intra-articular and topical, nasal orbuccal administration.

By the term “dosage unit” is meant a unitary, i.e. a single dose whichis capable of being administered to a patient, and which may be readilyhandled and packed, remaining as a physically and chemically stable unitdose comprising either the active material as such or a mixture of itwith solid or liquid pharmaceutical diluents or carriers.

The formulations may conveniently be presented in dosage unit form andmay be prepared by any of the methods well known in the art of pharmacy.All methods include the step of bringing the active ingredient intoassociation with the carrier which constitutes one or more accessoryingredients. In general, the formulations are prepared by uniformly andintimately bringing the active ingredient into association with a liquidcarrier or a finely divided solid carrier or both, and then, ifnecessary, shaping the product into the desired formulation.

Formulations of the present invention suitable for oral administrationmay be in the form of discrete units as capsules, sachets, tablets orlozenges, each containing a predetermined amount of the activeingredient; in the form of a powder or granules; in the form of asolution or a suspension in an aqueous liquid or non-aqueous liquid; orin the form of an oil-in-water emulsion or a water-in-oil emulsion. Theactive ingredient may also be administered in the form of a bolus,electuary or paste.

Formulations for rectal administration may be in the form of asuppository incorporating the active ingredient and a carrier, or in theform of an enema.

Formulations suitable for parenteral administration convenientlycomprise a sterile oily or aqueous preparation of the active ingredientwhich is preferably isotonic with the blood of the recipient.Transdermal formulations may be in the form of a plaster.

Formulations suitable for intra-articular or ophthalmic administrationmay be in the form of a sterile aqueous preparation of the activeingredient which may be in microcrystalline form, for example, in theform of an aqueous microcrystalline suspension. Liposomal formulationsor biodegradable polymer systems may also be used to present the activeingredient for both intra-articular and ophthalmic administration.

Formulations suitable for topical or ophthalmic administration includeliquid or semi-liquid preparations such as liniments, lotions, gels,applicants, oil-in-water or water-in-oil emulsions such as creams,ointments or pastes; or solutions or suspensions such as drops.

Formulations suitable for administration to the nose or buccal cavityinclude powder, self-propelling and spray formulations, such as aerosolsand atomizers.

In addition to the aforementioned ingredients, the formulations of thisinvention may include one or more additional ingredients, such asdiluents, binders, preservatives etc.

The compositions may further contain other therapeutically activecompounds usually applied in the treatment of the above mentionedpathological conditions, such as other immunosuppressants in thetreatment of immunological diseases, or steroids in the treatment ofdermatological diseases.

The present invention further concerns a method for treating patientssuffering from one of the above pathological conditions, said methodconsisting of administering to a patient in need of treatment aneffective amount of one or more compounds of formula I, alone or incombination with one or more other therapeutically active compoundsusually applied in the treatment of said pathological conditions. Thetreatment with the present compounds and/or with further therapeuticallyactive compounds may be simultaneous or with intervals.

In the systemic treatment daily doses from 0.001-2 μg per kilogram bodyweight, preferably from 0.002-0.3 μg/kg of mammal body weight, forexample 0.003-0.2 μg/kg of a compound of formula I are administered,typically corresponding to a daily dose for an adult human of from 0.2to 15 μg. In the topical treatment of dermatological disorders,ointments, creams or lotions containing from 0.1-500 μg/g, andpreferably from 0.1-100 μg/g, of a compound of formula I areadministered. For topical use in ophthalmology ointments, drops or gelscontaining from 0.1-500 μg/g, and preferably from 0.1-100 μg/g, of acompound of formula I are administered. The oral compositions areformulated, preferably as tablets, capsules, or drops, containing from0.05-50 μg, preferably from 0.1-25 μg, of a compound of formula I, perdosage unit.

The invention is further illustrated by the following GeneralProcedures, Preparations and Examples:

EXAMPLES General Procedures, Preparations and Examples

General:

THF was dried over 4A molecular sieves. Reactions were routinely rununder an argon atmosphere unless otherwise noted. In the standardwork-up procedure, the reaction mixture was poured into water andextracted three times with a suitable organic solvent. The organic layerwas washed with water and saturated sodium chloride solution, dried overanhydrous sodium sulfate, and concentrated in vacuo to give the product,which was purified by chromatography on silica gel (40-63 μm), bycrystallisation, or by distillation.

For ¹H nuclear magnetic resonance spectra (300 MHz) and ¹³C NMR (75.6MHz) chemical shift values (δ) (in ppm) are quoted, fordeuteriochloroform solutions, except where noted, relative to internaltetramethysilane (δ=0.00) or chloroform (δ=7.25) or deuteriochloroform(δ=76.81 for ¹³C NMR). The value for a multiplet, either defined(doublet (d), triplet (t), quartet (q)) or not (m) at the approximatemidpoint is given unless a range is quoted (s=singlet, b=broad).

General Procedures General Procedure 1 Synthesis of Ketones and 503 and504

To a solution/suspension of the appropriate silylated ethyl lactate,compound 501 or 502, (23.2 g, 0.1 mol) and N,O-dimethyl hydroxyl aminehydrochloride (12.2 g, 0.125 mol) in dry THF (800 ml) was added a 3.0molar solution of ethyl magnesium bromide in ether (167 ml, 0.5 mol),during one hour, while stirring and cooling to −5° C. Stirring wascontinued for 18 hours at 25° C., after which the reaction mixture washydrolyzed by pouring it into a solution of ammonium chloride (160 g) inwater (1.2 l). After work up with ether, the crude product was purifiedby chromatography (eluant: 0% to 1% ether in petroleum ether) to givethe desired compound.

Variation:

General Procedure 1a Synthesis of Ketones 509 and 510

To a solution/suspension of the appropriate R or S, methyl or ethyl,2-fluoropropionate (0.1 mol) and N,O-dimethylhydroxylamine hydrochloride(10.7 g, 0.11 mol) in dry THF (300 ml) was added a 3.0 molar solution ofethylmagnesium bromide in ether (107 ml, 0.32 mol), during one hour,while stirring and cooling to 0 to −5° C. Stirring was continued for 4hours at 0° C. and 16 hours at 25° C., after which the reaction mixturewas hydrolyzed by addition of 25% aqueous ammonium chloride (300 ml)with vigorous stirring. The temperature rose to about 35° C. and waskept there by means of a hot water bath; the pH was monitored with apH-meter (pH about 8). After 15 minutes the pH was adjusted to 3 with 4Naqueous hydrochloric acid and stirring was continued for 15 minutes. Theresulting two-phase mixture was separated and the aqueous phase wasextracted twice with ether (50 ml). The combined organic phases wereextracted twice with saturated sodium chloride solution (25 ml) anddried over anhydrous sodium sulfate. The solution of crude material wasconcentrated by distilling off the ether and some of the THF atatmospheric pressure through an an efficient still ({fraction (1/16)}″Dixon gauze rings packed in a 7 cm column). The residue (about 200 g)was fractionated through a 15 cm Vigreux-column connected to a dry-icecooled condenser and receiver. By gradually lowering the pressure to0.25 bar fractions consisting of THF and an increasing part of the titlecompound were collected (leaving a residue of higher boilingintermediate- and side-products). If desired, the first fractions may beconcentrated by redestillation. The pure title compounds have a b.p. ofabout 45° C./27 mbar; fractions mixed with THF were, however, usable inthe next synthetic step, when dried and the percentage of title compounddetermined by NMR.

General Procedure 2 Synthesis of Tertiary Propargylic Alcohols,Compounds II

A mixture of aluminium scales (0.45 g), mercury(II) chloride (12 mg) anddry THF (10 ml) was stirred for 20 minutes. A solution of propargylbromide (1.88 ml, 2.97 g, 25 mmol) in dry THF (5 ml) was added, withstirring, during 20 minutes, at 25°-30° C. Stirring was continued for 30minutes at 40° C. A solution of the appropriate ketone, Compound 503 or504, (5.0 g, 23 mmol) in dry THF (10 ml) was added, during 10 minutes,at 25° C.; the reaction mixture was stirred for a further 1½ hours at25° C. and then worked up with ether. The crude product was purified bychromatography, with 1% ether in petroleum ether as eluant, whichprocedure was repeated twice, with suitable combination of fractions, togive each of the two isomeric title compounds in a pure state.

Variation:

General Procedure 2a Synthesis of Fluorinated Tertiary PropargylicAlcohols, Compounds II

A 1.5 M solution of allenylmagnesium bromide in ether (13 ml, about 20mmol) (L. Brandsma, “Preparative Acetylenic Chemistry”, 2^(nd) Ed.,Elsevier, Amsterdam 1988, pp. 35-36) was cooled to −40° C., and a drysolution of the ketone 509 or 510 (16 mmol) in THF (20 ml) was addedwith stirring, during 30 minutes, followed by stirring for 10 minutes,while cooling to −30° C.-−40° C. The reaction mixture was poured into25% aqueous ammonium chloride (50 ml), while stirring and cooling inice. The aqueous phase was extracted with ether (20 ml) and the combinedorganic phases were dried with magnesium sulfate and concentrated invacuo to give the title compound as an oil. This product consisted ofthe two epimers in a ratio of about 3:1 and was used in the nextsynthetic step without further purification.

General Procedure 3 Synthesis of Tertiary Acetylenic Alcohols, CompoundsII

To a stirred suspension of lithium acetylide, ethylenediamine complex(5.3 g, 58 mmol) in dry THF (250 ml) was added a solution of theappropriate ketone, Compound 503 or 504, (12.5 g, 58 mmol) in dry THF(50 ml), during 20 minutes, at 35° C. Stirring was continued for 4 horsat 25° C., after which the reaction mixture was hydrolyzed by pouring itinto a solution of sodium chloride (780 g) in water (200 ml). After workup with ether, the crude product was purified by chromatography (eluant:0% to 2% ether in petroleum ether) to give the two isomeric titlecompounds as an only partially separable, but otherwise pure, mixture.This mixture was used as such in the next step, the removal of the silylprotection group, after which the two isomeric 25, 26-diols could beseparated by means of chromatography.

General Procedure 3a Synthesis of Tertiary Trimethylsilyl AcetylenicAlcohols, Compounds II

To a stirred solution of trimethylsilylacetylene (0.25 ml, 0.177 g, 1.8mmol) in dry THF (6 ml) was added a 1.6 molar solution of butyl lithiumin hexane (0.83 ml, 1.3 mmol), during 10 minutes, at −60° C. Stirringwas continued for 10 minutes at −60° C. and at 25° C. for 30 minutes.The resulting solution of lithium trimethylsilylacetylide was againcooled to −60° C. and a solution of the appropriate ketone, Compound 503or 504, (0.3 g, 1.4 mmol) in dry THF (1 ml), was added during 2 minutes.Stirring was continued for 3 hours at −60° C., after which the reactionmixture was hydrolyzed by pouring it into a solution of ammoniumchloride (2 g) in water (20 ml). After work up with ether, the crudeproduct was purified by chromatography (eluant: 0% to 1% ether inpetroleum ether) to give the two isomeric title compounds as aninseparable mixture. This was used as such in the next step, the removalof both silyl protection groups, after which the two isomeric25,26-diols could be separated by means of chromatography.

General Procedure 4 Deprotection of Silyl-Protected Compounds with TBAF

To a solution of the appropriate mono-silyl protected compound(s) (0.4mmol), or di-silyl protected compound(s) (0.2 mmol), or tetra-silylprotected compound(s) (0.1 mmol) in THF (10 ml) was added TBAFtrihydrate (0.32 g, 1.0 mmol), (i.e. 2.5 mol of TBAF for each molarequivalent of silyl protection group), and the mixture was heated toreflux for one hour with stirring. After addition of 1M sodium hydrogencarbonate (10 ml), the mixture was worked up with ether or ethylacetate. The residue was purified by chromatography to yield the desiredcompound(s).

General Procedure 5 Protection of 25,26-Diols II as Acetonides, Using2-Methoxypropene

A solution of an unprotected 25,26-diol, Compound II, (0.35 mmol),2-methoxypropene (50 mg, 0.7 mmol) and p-toluenesulfonic acid (1 mg) indry DMF (2 ml), containing 3 Å molecular sieves ({fraction (1/16)}″rods, about 0.1 g) was heated to 70° C., with stirring, for 30° minutes.After work up with ether, the crude product was purified bychromatography with 5% ether in petroleum ether as eluant to give thepure acetonide, Compound II.

General Procedure 5a Protection of 25,26-Diols II as Acetonides, Using2,2-Dimethoxypropane

A solution of an unprotected 25,26-diol, Compound II, (2.0 mmol),2,2-dimethoxy-propane (2 ml, 1.7 g, 16 mmol) and p-toluene sulfonic acid(38 mg) in dry acetone (10 ml) was stirred at 25° C. 2 hours.Triethylamine (1 ml) was added and the reaction mixture was concentratedto about 2 ml, using a short (about 3 cm) Vigreux column, at about b.p.30° C./0.4 bar. Water (2 ml) and 1M sodium hydrogen carbonate (1 ml)were added, and the mixture was extracted with ether (3×8 ml). Thecombined ether extracts were dried with magnesium sulfate, andconcentrated to about 0.5 ml (as above). The crude product was purifiedby chromatography with 5% ether in petroleum ether as eluant. Afterconcentration (as above) of the appropriate combined fractions the pureacetonide, Compound II, was obtained.

General Procedure 6 Protection of 25-Monohydroxy Compounds II byTrimethylsilylation

Trimethylchlorosilane (1.08 g, 10 mmol) was added during 10 minutes, at0° C., to a stirred solution of a25-hydroxy-26-tert-butyldimethylsilyloxy- (or 26-fluoro-) compound II(5.0 mmol), triethylamine (1.51 g, 15 mmol) and DMAP (5 mg) in drydichloromethane (10 ml). Stirring was continued for 48 hours at 25° C.After work up (with ether) the crude product was purified bychromatography with petroleum ether as eluant to give the pure25-trimethylsilyloxy-26-tert-butyldimethylsilyloxy- (or 26-fluoro-)compound II.

General Procedure 7 Synthesis of Compounds III from Compound 1 and SideChain Building Block II

To a solution of the appropriate side chain building block, Compound II,(3.0 mmol) in dry HF (5 ml), cooled to −78° C. and stirred under argon,was added dropwise, during 2 minutes, a solution of n-butyl lithium (1.6M in hexane; 1.5 ml). Stirring was continued at −78° C. for 15 minutesand at 20° C. for another 15 minutes. The mixture was again cooled to−78° C., and a solution of the aldehyde, compound 1, (1.5 mmol) in dryTHF (5 ml) was added dropwise during 4 minutes; after that stirring wascontinued at −78° C. for 30 minutes. The reaction mixture was worked up(ether) to yield a crude product containing the isomeric 22-hydroxycompounds A (less polar) and B (more polar). These were separated bychromatography (mixture of ethyl acetate and petroleum ether as eluant)to yield the pure compounds III.

Variation:

General Procedure 7a

The procedure of G.P. 7 was followed, except that 1.87 ml of n-butyllithium (1.6M) and 3 mmol of compound 1 was used.

General Procedure 8 Photoisomerisation of Compound III to Compound IV

A solution of the appropriate compound III (0.28 mmol), anthracene (0.1g) and triethylamine (0.20 ml, 1.4 mmol) in dichloromethane (16 ml) in a25 ml round-bottomed Pyrex flask was irradiated at about 10° C. withUV-light from a high pressure ultraviolet lamp, type TQ760Z2 (Hanau), at700 W, for 30 minutes (15 minutes at 0.08 mmol scale) while stirring.The reaction mixture was evaporated in vacuo, and the residue wastreated with petroleum ether (2×2 ml) and filtered. The filtrate wasconcentrated and purified by chromatography to afford the desiredcompound IV.

Variation:

General Procedure 8a

A solution of the appropriate compound III (0.13 mmol),9-acetylanthracene (23 mg) and triethylamine (0.10 ml, 0.7 mmol) intoluene (5 ml) in a 10 ml round-bottomed Pyrex tube was irradiated atabout 10° C. with UV-light from a high pressure ultraviolet lamp, typeTQ150Z2 (Hanau) (150 W) for 60 minutes. The reaction mixture was cooledto −20° C. and filtered. The filtrate was evaporated in vacuo, and theresidue was purified by chromatography to afford the desired compoundIV, together with some 9-acetyl-anthracene which was removed after thefinal (deprotection) step.

General Procedure 9 Alkylation of C-22-Hydroxy-Compound III or IV

To a solution of the appropriate 22-hydroxy compound (R═H) (0.5 mmol) indry THF (5 ml) was added, while stirring at 20° C. under argon, a 20%suspension of potassium hydride in mineral oil (0.2 ml) followed by analkylating agent, RZ (1.5 mmol) and, finally, during 5 minutes, asolution of 18-Crown-6 (0.13 g) in dry THF (2 ml). Stirring at 25° C.was continued for two hours, after which the reaction mixture was workedup (ether). The crude product was purified by chromatography (mixture ofether and petroleum ether as eluant) to yield the desired alkoxycompound III or IV.

General Procedure 10 Synthesis of Compound I by Deprotection of CompoundIV with HF

To a stirred solution of the appropriate Compound IV (0.25 mmol) inethyl acetate (3 ml) was added acetonitrile (6 ml) followed by a 5%solution of hydrofluoric acid in acetonitrile-H₂O 7:1 (4 ml). Afterstirring for 2 hours at 25° C. ethyl acetate (40 ml) and 1M sodiumhydrogen carbonate (20 ml) was added, and the reaction mixture wasworked up (ethyl acetate). The residue was purified by chromatography togive the desired compound I.

General Procedure 11 Synthesis of Lactic Ester Mesylates

A solution of methanesulfonyl chloride (18.6 ml; 27.5 g; 0.24 mol) intert-butyl methyl ether (100 ml) was added during one hour to a stirredsolution of the appropriate R or S, methyl or ethyl, lactate (0.20 mol),triethylamine (28.3 g; 0.28 mol) and DMAP (0.24 g; 0.002 mol) intert-butyl methyl ether (200 ml), while cooling in an ice bath. Stirringwas continued for ½ hour in the ice bath a followed by 2 hours at 25° C.The reaction mixture was again cooled in an ice bath, and water (250 ml)was added slowly, keeping the temperature below 10° C. After stirringfor 20 minutes more, the phases were separated, and the aqueous phasewas extracted twice with ether (100 ml). The combined organic phaseswere extracted with: 1 N sulfuric acid (100 ml), water (100 ml), 1 Msodium hydrogen carbonate (100 ml), water (2×100 ml) and saturatedaqueous sodium chloride (100 ml). After drying with sodium sulfate thesolution was concentrated (at 35° C. and 0.2 bar), and the residuefractionated in vacuo through a 45 cm Podbielniak-type column to givethe desired title compound.

General Procedure 12 Synthesis of R or S, Methyl or Ethyl,2-Fluoropropionate

A solution of the appropriate lactic ester mesylate (0.125 mol) wasadded, during about one hour, to a solution/suspension of (freeze-dried)potassium fluoride (29 g; 0.5 mol) in formamide (70 ml) which wasstirred and heated in an oil bath at 60° C., in a vacuum of about 27mbar. The flask was equipped with a Claisen-type side arm which wasconnected to a dry-ice cooled and a dry-ice cooled receiver, in whichwas condensed the 2-fluoropropionic acid ester that was destilledcontinuously from the reaction. Stirring at 60° C. and 20 mm Hg wascontinued until no more distillate was condensed, this took 4-5 hours.The distillate (which contained some water) was diluted with ether (40ml), dried with magnesium sulfate and purified by distillation in asetup similar to the one used for the above preparation, keeping thebath temperature fairly constant at 50-60° C. and gradually lowering thepressure, until the pure title compound was collected in the receiver.

Preparation 1

Compound 502

A solution of (+)-ethyl D-lactate (R-isomer, unnatural) (5.2 g, 44mmol), imidazole (13.6 g) and tert-butyl dimethylsilylchloride (15 g) indry DMF (50 ml) was stirred at 25° C. for 1 hour. Ethyl acetate (200 ml)was added and the organic solution was extracted with water (2×100 ml),3M CaCl₂ (2×100 ml), water (100 ml) and 35% NaCl (100 ml), dried withsodium sulfate and concentrated in vacuo. The crude product was purifiedby chromatography with 1% ether in petroleum ether as eluant to giveCompound 502 as an oil.

[α]_(D) ²⁰+30.5° (c 2.14, CHCl₃)

¹³C NMR δ 173.9, 68.3, 60.5, 25.5, 21.1, 18.1, 14.0, −5.1, −5.5

Preparation 2

Compound 503

Method: General Procedure 1

Staring material: Ethyl (S)-(−)-O-tert-butyldimethylsilyllactate(Compound 501)

Chromatography eluant: 0% to 5% ether in petroleum ether

[α]_(D) ²⁰−10.8° (c 2.17, CHCl₃)

¹³C NMR δ 214.7, 74.6, 29.9, 25.5, 20.8, 17.8, 7.1, −4.9, −5.3

Preparation 3

Compound 504

Method: General Procedure 1

Starting material: Compound 502

Chromatography eluant: 0% to 5% ether in petroleum ether.

¹³C NMR δ 214.7, 74.6, 29.9, 25.5, 20.8, 17.8, 7.1, −4.9, −5.3

Preparation 4

Compound 201(+Compound 205)

Method: General Procedure 2

Starting material; Compound 503

Chromatography eluant: 1% ether in petroleum ether.

¹³C NMR δ ¹H NMR d 81.1, 75.6, 71.3, 70.0, 26.5, 25.8, 25.6, 17.8, 17.1,7.4 −4.3, −5.2

Preparation 5

Compound 202

Method: General Procedure 4

Starting material: Compound 201

Chromatography eluant: 25% to 50% ether in petroleum ether.

¹³C NMR δ 80.7, 75.5, 71.2, 71.1, 27.1, 26.0, 17.1, 7.6

Preparation 6

Compound 203

Method: General Procedure 5

Starting material: Compound 202

Chromatography eluant: 5% ether in petroleum ether.

¹³C NMR δ 106.8, 82.4, 80.2, 78.2, 70.8, 28.3, 26.7, 25.6, 25.0, 14.6,7.2

Preparation 7

Compound 204

Method: General Procedure 6

Starting material: Compound 201

Chromatography eluant: petroleum ether.

[α]_(D) ²⁰−4.4° (c 1.8, CHCl₃)

¹³C NMR δ 82.2, 80.1, 71.4, 70.3, 28.8, 25.8, 24.9, 17.9, 17.3, 7.5,2.4, −4.4, −4.9

Preparation 8

Compound 205 (+Compound 201)

Method: General Procedure 2

Starting material: Compound 503

Chromatography eluant: 1% ether in petroleum ether.

¹³C NMR δ 80.8, 75.1, 72.3, 70.2, 27.8, 25.6, 24.4, 17.8, 17.6, 7.4,−4.3, −5.2

Preparation 9

Compound 206

Method: General Procedure 4

Starting material: Compound 205

Chromatography eluant: 25 to 50% ether in petroleum ether.

¹³C NMR δ 80.8, 75.2, 71.6, 71.1, 29.1, 24.7, 17.0, 7.4

Preparation 10

Compound 207

Method: General Procedure 5

Starting material: Compound 206

Chromatography eluant: 5% ether in petroleum ether.

¹³C NMR δ 106.9, 83.1, 81.0, 76.8, 70.1, 28.4, 28.2, 26.5, 24.5, 14.5,8.1

Preparation 11

Compound 208

Method: General Procedure 6

Starting material: Compound 205

Chromatography eluant: petroleum ether.

[α]_(D) ²⁰+6.9° (c 1.6, CHCl₃)

¹³C NMR δ 82.0, 79.8, 72.7, 70.5, 27.9, 27.0, 25.8, 25.7, 17.6, 8.3,2.5, −4.4, −5.0

Preparation 12

Compound 209 (+Compound 211)

Method: General Procedure 2

Starting material: Compound 504

Chromatography eluant: 1-2% ether in petroleum ether.

¹³C NMR δ 81.1, 75.6, 71.3, 70.0, 26.5, 25.8, 25.6, 17.8, 17.1, 7.4,−4.3, −5.2

Preparation 13

Compound 210

Method: General Procedure 6

Starting material: Compound 209

Chromatography eluant: petroleum ether.

[α]_(D) ²⁰+4.1° (c 2.1, CHCl₃)

¹³C NMR δ 82.2, 80.1, 71.4, 70.3, 28.8, 25.8, 25.0, 17.9, 17.3, 7.5,2.4, −4.4, −4.9

Preparation 14

Compound 211 (+Compound 209)

Method: General Procedure 2

Starting material: Compound 504

Chromatography eluant: 1% to 2% ether in petroleum ether.

¹³C NMR δ 80.8, 75.1, 72.3, 70.2, 27.8, 25.6, 24.4, 17.8, 17.6, 7.4,−4.3, −5.2

Preparation 15

Compound 212

Method: General Procedure 6

Starting material: Compound 211

Chromatography eluant: petroleum ether.

[α]_(D) ²⁰−7.0° (c 1.2, CHCl₃)

¹³C NMR δ 82.0, 79.8, 72.8, 70.5, 27.9, 27.0, 25.7, 17.8, 17.6, 8.3,2.5, −4.4, −5.0

Preparation 16

Compound 213+Compound 217

Method: General Procedure 3

Starting material: Compound 503

Chromatography eluant: 0% to 2% ether in petroleum ether.

Compound 213

¹H NMR δ 3.70 (q,1H), 2.51 (s,1H), 1.80 (m,1H), 1.64 (m,1H), 1.25(d,3H), 0.98 (t,3H), 0.88 (s,9H), 0.20 (s,6H)

Compound 217

¹³C NMR δ 83.9, 74.7, 73.3, 73.1, 31.0, 25.6, 25.5, 18.6, 8.1, −4.3,−5.1

Preparation 17

Compound 214+Compound 218

Method: General Procedure 3a

Starting material: Compound 503

Chromatography eluant: 0% to 1% ether in petroleum ether.

Compound 214

¹³C NMR δ 105.9, 89.2, 74.6, 73.5, 30.9, 25.7, 18.7, 17.2, 8.3, −0.3,−4.5, −4.9

Compound 218

¹³C NMR δ 105.9, 89.6, 75.1, 73.4, 30.9, 25.7, 25.6, 18.7, 8.3, −0.3,−4.2, −5.0

Preparation 18

Compound 215+Compound 219

Method: General Procedure 4

Starting material: Compound 213+Compound 217 or Compound 214+Compound218

Chromatography eluant: a) 40% ether in petroleum ether, b) 33% ether inpetroleum ether

Compound 215

¹H NMR δ 3.84 (1H,q), 2.49 (1H,s), 1.74 (1H,m), 1.61 (1H,m), 1.27(3H,d), 1.09 (3H,t)

Compound 219

Crystallised from ether/n-heptane, m.p. 52°-53°.

¹H NMR δ 3.65 (q,1H), 2.46 (s,1H), 1.65 (m,2H), 1.31 (d,3H), 1.08 (t,3H)

Preparation 19

Compound 216

Method: General Procedure 5a

Starting material: Compound 219

Chromatography eluant: 5% ether in petroleum ether.

¹³C NMR δ 108.6, 83.9, 79.3, 79.2, 72.8, 29.3, 28.3, 26.0, 13.6, 8.2

Preparation 20

Compound 220

Method: General Procedure 5a

Starting material: Compound 215

Chromatography eluant: 5% ether in petroleum ether.

¹³C NMR δ 108.6, 82.8, 81.3, 78.4, 75.1, 31.3, 27.6, 27.1, 15.8, 8.8

Preparation 21

Compound 301+Compound 302

Method: General Procedure 7

Starting material: Compound 204

Chromatography eluant: 0% to 10% ether in petroleum ether.

Compound 301

¹³C NMR δ 153.4, 142.8, 135.3, 121.5, 116.4, 106.5, 83.2, 82.9, 80.3,71.6, 70.1, 67.0, 64.6, 56.0, 51.4, 45.5, 43.8, 41.0, 39.6, 36.4, 29.0,28.7, 26.4, 25.8, 25.7, 25.6, 25.2, 23.2, 21.9, 18.1, 17.9, 17.9, 17.3,13.0, 12.4, 7.6, 2.5, −4.3, −5.0, −5.1

Compound 302

¹³C NMR δ 152.5, 141.9, 134.3, 120.5, 115.4, 105.5, 82.3, 82.0, 79.3,70.7, 69.1, 66.0, 64.7, 55.1, 51.1, 44.6, 42.8, 41.5, 39.2, 35.5, 28.0,27.7, 26.0, 24.9, 24.7, 24.6, 24.4, 22.3, 21.0, 17.1, 17.0, 16.9, 16.4,12.4, 10.8, 6.6, 1.5, −5.3, −5.8, −5.9

Preparation 22

Compound 401

Method: General Procedure 8

Starting material: Compound 301

Chromatography eluant: 0% to 2% ether in petroleum ether.

¹³C NMR δ 148.2, 140.4, 135.0, 122.9, 117.9, 111.0, 83.1, 82.9, 80.3,71.8, 71.6, 67.3, 64.6, 55.9, 51.4, 45.8, 45.3, 44.6, 41.0, 39.7, 29.0,28.6, 26.5, 25.8, 25.7, 25.6, 25.2, 23.2, 21.8, 18.0, 17.9, 17.3, 13.0,12.3, 7.6, 2.5, −4.3, −4.9, −5.0, −5.3

Preparation 23

Compound 402

Method: General Procedure 9

Starting material: Compound 401

Alkylating agent: Methyl iodide

Chromatography eluant: 0% to 1% ether in petroleum ether.

¹³C NMR δ 148.2, 140.6, 134.9, 122.9, 117.8, 111.0, 83.9, 80.8, 80.4,73.6, 71.8, 71.7, 67.4, 56.0, 55.8, 51.4, 45.8, 45.3, 44.6, 40.4, 39.4,29.1, 28.7, 26.5, 25.9, 25.7, 25.6, 25.3, 23.3, 21.8, 19.2, 18.0, 18.0,17.4, 13.8, 12.4, 7.6, 2.5, −4.4, −4.9, −5.0, −5.3

Preparation 24

Compound 403

Method: General Procedure 9

Starting material: Compound 401

Alkylating agent: Ethyl bromide

Chromatography eluant: 0% to 1% ether in petroleum ether.

¹³C NMR δ 148.2, 140.7, 134.8, 123.0, 117.7, 111.0, 83.3, 81.4, 80.4,72.0, 71.8, 71.7, 67.4, 63.8, 55.9, 51.4, 45.8, 45.4, 44.6, 40.3, 39.3,29.1, 28.7, 26.4, 25.9, 25.7, 25.6, 25.3, 23.3, 21.8, 18.1, 18.0, 17.4,15.0, 13.9, 12.4, 7.6, 2.5, −4.4, −4.9, −5.0, −5.3

Preparation 25

Compound 404

Method: General Procedure 9

Starting material: Compound 401

Alkylating agent: Propyl bromide

Chromatography eluant: 0% to 1% ether in petroleum ether.

¹³C NMR δ 148.2, 140.8, 134.8, 123.0, 117.7, 111.0, 83.3, 81.5, 80.5,72.1, 71.8, 71.7, 70.1, 67.4, 55.9, 51.4, 45.8, 45.4, 44.6, 40.4, 39.5,29.1, 28.7, 26.3, 25.9, 25.7, 25.6, 25.3, 23.3, 22.9, 21.8, 18.0, 18.0,17.3, 13.9, 12.4, 10.7, 7.6, 2.5, −4.4, −4.9, −5.0, −5.3

Preparation 26

Compound 405

Method: General Procedure 9

Starting material: Compound 401

Alkylating agent: Benzyl bromide

Chromatography eluant: 0% to 1% ether in petroleum ether.

¹³C NMR δ 148.2, 140.7, 138.5, 134.9, 128.2, 128.0, 127.6, 127.4, 127.1,122.9, 117.7, 111.0, 84.1, 81.0, 80.4, 71.8, 71.7, 70.1, 67.4, 55.8,51.4, 45.8, 45.4, 44.6, 40.4, 39.4, 29.2, 28.7, 26.3, 25.9, 25.7, 25.6,25.3, 23.2, 21.8, 18.0, 18.0, 17.4, 14.0, 12.4, 7.7, 2.5, −4.4, −4.8,−4.9, −5.0, −5.3

Preparation 27

Compound 303

Method: General Procedure 7

Starting material: Compound 208

Chromatography eluant: 0% to 5% ether in petroleum ether.

¹³C NMR δ 153.4, 142.8, 135.3, 121.5, 116.4, 106.5, 83.1, 82.9, 80.0,77.0, 72.9, 70.1, 67.0, 64.6, 56.0, 51.4, 45.5, 43.8, 41.0, 39.6, 36.4,28.7, 28.0, 27.2, 26.4, 25.8, 25.7, 25.7, 25.6, 23.2, 21.9, 18.1, 17.9,17.7, 13.0, 12.4, 8.4, 2.5, −4.4, −5.0, −5.1, −5.1

Preparation 28

Compound 406

Method: General Procedure 8

Starting material: Compound 303

Chromatography eluant: 0% to 5% ether in petroleum ether.

¹³C NMR δ 148.2, 140.5, 135.0, 122.9, 117.9, 111.0, 83.1, 82.9, 80.0,72.9, 71.8, 67.3, 64.6, 55.9, 51.4, 45.8, 45.3, 44.6, 41.0, 39.7, 28.6,28.0, 27.2, 26.5, 25.8, 25.7, 25.7, 25.6, 23.2, 21.8, 18.0, 17.9, 17.9,17.7, 13.0, 12.3, 8.4, 2.5, −4.4, −4.9, −4.9, −5.0, −5.3

Preparation 29

Compound 407

Method: General Procedure 9

Starting material: Compound 406

Alkylating agent: Ethyl bromide

Chromatography eluant: 0% to 2% ether in petroleum ether.

13C NMR δ 148.2, 140.8, 134.8, 123.0, 117.7, 111.0, 83.1, 81.4, 80.1,72.9, 71.9, 71.8, 67.4, 63.8, 55.9, 51.4, 45.8, 45.4, 44.6, 40.4, 39.4,28.7, 28.0, 27.2, 26.5, 25.8, 25.7, 25.6, 23.3, 21.8, 18.0, 18.0, 17.9,17.7, 15.0, 13.9, 12.4, 8.3, 2.4, −4.4, −4.9, −4.9, −5.0, −5.3

Preparation 30

Compound 304

Method: General Procedure 7

Starting material: Compound 210

Chromatography eluant: 5% ether in petroleum ether.

¹³C NMR δ 153.4, 142.8, 135.3, 121.5, 116.4, 106.5, 83.2, 82.9, 80.3,71.6, 70.1, 67.0, 64.6, 56.0, 51.4, 45.5, 43.8, 41.0, 39.6, 36.4, 28.9,28.7, 26.4, 25.8, 25.7, 25.6, 25.2, 23.2, 21.9, 18.1, 18.0, 17.9, 17.3,13.0, 12.4, 7.6, 2.5, −4.3, −4.9, −5.0, −5.1, −5.1

Preparation 31

Compound 408

Method: General Procedure 8

Starting material: Compound 304

Chromatography eluant: 5% ether in petroleum ether.

¹³C NMR δ 148.2, 140.4, 135.0, 122.9, 117.9, 111.0, 83.1, 83.0, 80.3,71.8, 71.6, 67.3, 64.9, 55.9, 51.4, 45.8, 45.3, 44.6, 41.0, 39.7, 28.9,28.6, 26.5, 25.8, 25.7, 25.7, 25.6, 25.2, 23.2, 21.8, 18.0, 17.9, 17.3,13.0, 12.3, 7.6, 2.5, −4.3, −4.9, −5.0, −5.3

Preparation 32

Compound 409

Method: General Procedure 9

Starting material: Compound 408

Alkylating agent: Ethyl bromide

Chromatography eluant: 1% ether in petroleum ether.

¹³C NMR δ 148.2, 140.7, 134.8, 123.0, 117.7, 111.0, 83.4, 81.4, 80.4,72.0, 71.8, 71.7, 67.8, 67.4, 63.8, 55.9, 51.3, 45.8, 45.4, 44.6, 40.3,39.3, 29.0, 28.7, 26.3, 25.8, 25.7, 25.6, 25.4, 25.3, 23.3, 21.8, 18.0,18.0, 17.4, 15.0, 14.0, 12.4, 7.6, 2.5, −4.4, −4.9 −5.0, −5.3

Preparation 33

Compound 305

Method: General Procedure 7

Starting material: Compound 212

Chromatography eluant: 3% ether in petroleum ether.

¹³C NMR δ 263.2, 153.4, 142.8, 135.3, 121.5, 116.4, 106.5, 82.9, 80.0,72.9, 70.1, 67.0, 64.7, 56.0, 51.4, 45.5, 43.8, 41.0, 39.6, 36.4, 28.7,28.0, 27.2, 26.4, 25.7, 25.7, 25.6, 23.2, 21.9, 18.1, 17.9, 17.6, 13.0,12.4, 8.4, 2.5, −4.4, −5.0, −5.1

Preparation 34

Compound 410

Method: General Procedure 8

Starting material: Compound 305

Chromatography eluant: 3% ether in petroleum ether.

¹³C NMR δ 148.2, 140.5, 135.0, 122.9, 117.9, 111.0, 83.1, 82.9, 80.0,72.9, 71.8, 67.3, 64.7, 55.9, 51.4, 45.8, 45.3, 44.6, 41.0, 39.7, 28.7,28.0, 27.2, 26.5, 25.7, 25.7, 25.6, 23.2, 21.8, 18.1, 18.0, 17.9, 17.6,13.0, 12.3, 8.4, 2.5, −4.4, −4.9, −4.9, −5.0, −5.3

Preparation 35

Compound 411

Method: General Procedure 9

Starting material: Compound 410

Chromatography eluant: 1% ether in petroleum ether.

¹³C NMR δ 148.2, 140.8, 134.8, 123.0, 117.7, 111.0, 83.0, 81.4, 80.1,72.9, 72.0, 71.8, 67.4, 63.8, 55.9, 51.4, 45.8, 45.4, 44.7, 40.3, 39.3,28.7, 28.0, 27.2, 26.4, 25.8, 25.7, 25.6, 23.3, 21.8, 18.0, 18.0, 17.9,17.6, 15.0, 14.0, 12.4, 8.4, 2.5, −4.4, −4.9, −4.9, −5.0, −5.3

Preparation 36

Compound 306

Method: General Procedure 7a

Starting material: Compound 216

Chromatography eluant: 0% to 20% ether in petroleum ether.

¹³C NMR δ 153.4, 142.7, 135.4, 121.5, 116.4, 108.1, 106.5, 85.6, 84.8,79.3, 78.9, 70.1, 67.0, 64.4, 55.9, 51.2, 45.5, 43.8, 40.6, 39.5, 36.4,28.7, 28.2, 28.0, 26.2, 25.9, 25.7, 25.6, 23.2, 21.9, 18.1, 17.9, 13.5,13.1, 12.4, 8.2, −5.0, −5.1, −5.1

Preparation 37

Compound 412

Method: General Procedure 8

Starting material: Compound 306

Chromatography eluant: 10% ether in petroleum ether.

¹³C NMR δ 148.1, 140.3, 135.0, 122.9, 117.9, 111.0, 108.1, 85.6, 84.7,79.3, 78.9, 71.9, 67.3, 64.4, 55.8, 51.2, 45.8, 45.3, 44.6, 40.6, 39.5,28.6, 28.2, 28.0, 26.2, 25.9, 25.7, 25.6, 23.1, 21.8, 18.0, 17.9, 13.5,13.1, 12.4, 8.2, −4.9, −5.0, −5.3

Preparation 38

Compound 413

Method: General Procedure 9

Starting material: Compound 412

Alkylating agent: Ethyl bromide

Chromatography eluant: 0% to 2% ether in petroleum ether.

¹H NMR δ 6.22 (d,1H), 6.00 (d,1H), 5.17 (d,1H), 4.85 (d,1H), 4.36(m,1H), 4.30 (q,1H), 4.20 (d,1H), 4.16 (m,1H), 3.72 (m,1H), 3.30 (m,1H),2.82 (dd,1H), 2.43 (dd,1H), 2.20 (dd,1H), 1.99 (t,1H), 2.0-0.8 (m,15H),1.44 (s,3H), 1.40 (s,3H), 1.26 (d,3H), 1.18 (t,3H), 1.07 (t,3H), 1.00(d,3H), 0.87 (s,18H), 0.51 (s,3H), 0.05 (s,12H)

Preparation 39

Compound 307

Method: General Procedure 7a

Starting material: Compound 220

Chromatography eluant: 0% to 20% ether in petroleum ether.

¹³C NMR δ 153.4, 142.7, 135.3, 121.5, 116.4, 108.2, 106.5, 87.9, 83.8,81.2, 78.4, 70.1, 67.0, 64.4, 55.9, 51.3, 45.4, 43.8, 40.7, 39.5, 36.4,31.3, 28.7, 27.6, 26.8, 26.3, 25.7, 25.6, 23.2, 21.9, 18.1, 17.9, 15.8,13.1, 12.4, 8.7, −5.0, −5.1, −5.1

Preparation 40

Compound 414

Method: General Procedure 8

Starting material: Compound 307

No Chromatography: Crude product used in the next step

¹³C NMR δ 6.22 (1H,d), 6.01 (1H,d), 5.17 (1H,d), 4.85 (1H,d), 4.67(1H,s), 4.36 (1H,m), 4.17 (1H,m), 3.85 (1H,q), 2.82 (1H,dd), 2.43(1H,dd), 2.21 (1H,dd), 1.89 (1H,t), 1.85-1.00 (13H,m), 1.68 (2H,q), 1.51(3H,s), 1.35 (3H,d), 1.33 (3H,s), 1.06 (3H,t), 1.04 (3H,d), 0.87(18H,s), 0.53 (3H,s), 0.05 (12H,s)

Preparation 41

Compound 415

Method: General Procedure 9

Starting material: Compound 414

Alkylating agent: Ethyl bromide

Chromatography eluant: 0% to 2% ether in petroleum ether.

¹³C NMR δ 148.1, 140.7, 134.9, 122.9, 117.8, 111.0, 108.1, 86.5, 84.2,81.3, 78.5, 71.9, 71.8, 67.3, 63.9, 55.9, 51.4, 45.9, 44.6, 40.2, 39.3,31.4, 28.7, 27.5, 26.9, 26.3, 25.7, 25.6, 23.3, 21.8, 18.0, 18.0, 15.8,15.0, 14.0, 12.4, 8.8, −4.9, −5.0, −5.3

Preparation 42

Compound 505

Method: General Procedure 11

Starting material: Methyl (R)-(+)-lactate

Purification: Distillation; b.p. 94° C./1.3 mbar; [α]_(D) ²⁰+54.4° (c2.29, CHCl₃)

¹³C NMR δ 170.0, 74.1, 52.8, 39.1, 18.4

Preparation 43

Compound 506

Method: General Procedure 11

Starting material: Ethyl (S)-(−)-lactate

Purification: Distillation; b.p. 98° C./1.5 mbar; [α]_(D) ²⁰−53.1° (c2.03, CHCl₃), (litt.:

Breitschuh, R. et al., Synthesis, 1992, 1170: [α]_(D) ²⁰−54.6° (c 4.36.CHCl₃))

Preparation 44

Compound 507

Method: General Procedure 12

Starting material: Compound 505

Purification: Distillation; b.p. 39° C./53 mbar; [α]_(D) ²⁰−2.8° (c2.21, CHCl₃)

¹H NMR δ 5.02 (dq,1H), 3.80 (s,3H), 1.58 (dd,3H)

Preparation 45

Compound 508

Method: General Procedure 12

Starting material: Compound 506

Purification: Distillation; b.p. 34° c./27 mbar; [α]_(D) ²⁰+3.8° (c2.32, CHCl₃)

¹³C NMR δ 170.5, 85.7, 61.5, 18.3, 14.1

Preparation 46

Compound 509

Method: General Procedure 1a

Starting material: Compound 507

Purification: Distillation; b.p. about 45° C./27 mbar; [α]_(D) ²⁰−51.6°(approx.) (c 2.1, CHCl₃:THF 4:1)

¹H NMR δ 4.88 (dq, 1H); 2.64 (m, 2H); 1.47 (dd, 3H); 1.08 (t, 3H)

Preparation 47

Compound 510

Method: General Procedure 1a

Starting material: Compound 508

Purification: Distillation; b.p. about 45° C./27 mbar; [α]_(D) ²⁰+43.3°(approx.) (c 2.0, CHCl₃:THF 7:3)

¹³C NMR δ 210.8, 92.6, 30.7, 17.7, 6.8

Preparation 48

Compound 221

Method: General Procedure 2a

Purification: None

Starting material: Compound 509

¹³C NMR δ

Major 93.4, 79.6, 74.3, 71.3, 27.8, 25.0, 14.8, 7.4

Minor 93.0, 80.0, 74.6, 71.0, 27.0, 25.4, 14.7, 7.2

Preparation 49

Compound 222

Method: General Procedure 6

Starting material: Compound 221

Chromatography eluant: petroleum ether

Preparation 50

Compound 223

Method: General Procedure 2a

Starting material: Compound 210

Purification: None

¹³C NMR δ

Major: 93.4, 79.7, 74.3, 71.4, 27.8, 25.0, 14.8, 7.4

Minor: 93.0, 80.0, 74.8, 71.0, 27.0, 25.4, 14.7, 7.2

Preparation 51

Compound 224

Method: General Procedure 6

Starting material: Compound 223

Chromatography eluant: petroleum ether.

¹³C NMR δ

Major: 93.2, 80.4, 78.3, 70.5, 28.1, 25.9, 14.9, 7.7, 2.1

Minor: 92.5, 80.6, 78.2, 70.3, 27.9, 25.1, 14.6, 7.2, 2.1

Preparation 52

Compound 308

Method: General Procedure 7

Starting materials: Compound 1 and Compound 222

Chromatography eluant: 0% to 20% ether in petroleum ether

Preparation 53

Compound 416

Method: General Procedure 8a

Starting material: Compound 308

Chromatography eluant: 50% dichloromethane in petroleum ether

Preparation 54

Compound 417

Method: General Procedure 9

Starting material: Compound 416

Alkylating agent: Ethyl bromide

Chromatography eluant: 0% to 10% ether in petroleum ether

Preparation 55

Compound 309

Method: General Procedure 7

Starting materials: Compound 1 and Compound 224

Chromatography eluant: 0% to 20% ether in petroleum ether.

¹³C NMR δ 153.4, 142.7, 135.4, 121.5, 116.4, 106.5, 93.3, 83.1, 81.5,78.5, 70.1, 67.0, 64.6, 55.9, 51.4, 45.5, 43.8, 41.2, 40.9, 39.6, 36.4,28.9, 28.7, 28.3, 27.5, 26.3, 25.7, 25.6, 23.2, 22.4, 21.9, 20.2, 19.2,18.1, 17.9, 15.1, 14.8, 14.1, 13.0, 12.4, 11.2, 7.8, 2.2, −5.0, −5.1,−5.1

Preparation 56

Compound 418

Method: General Procedure 8a

Starting material: Compound 309

Chromatography eluant: 50% dichloromethane in petroleum ether.

¹H NMR δ 6.22 (d,1H), 6.02 (d,1H), 5.17 (d,1H), 4.85 (d,1H), 4.62(dq,1H), 4.60 (m,1H), 4.37 (m,1H), 4.18 (m,1H), 2.83 (d,1H, 2.50-0.80(m,24H), 1.31 (dd,3H), 1.01 (d,3H), 0.85 (s,18H), 0.83 (d,3H), 0.12(s,9H), 0.06 (d,12H)

Preparation 57

Compound 419

Method: General Procedure 9

Starting material: Compound 418

Alkylating agent: Ethyl bromide

Chromatography eluant: 0% to 10% ether in petroleum ether.

¹³C NMR δ 148.1, 140.6, 134.9, 122.9, 117.8, 111.0, 93.4, 82.6, 80.5,74.3, 71.9, 67.3, 64.0, 55.8, 51.3, 45.9, 45.4, 40.1, 39.3, 28.7, 27.8,25.7, 25.6, 25.0, 23.3, 21.8, 18.0, 17.9, 14.7, 14.0, 12.4, 7.2, 2.2,−4.9, −5.0, −5.3

EXAMPLES Example 1 1(S),3(R)-Dihydroxy-20(R)-(5-Ethyl-1(S),5(S),6(S)-Trihydroxy-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z), 7(E), 10(19)-Triene

Compound 101

Method: General Procedure 4

Starting material: Compound 401

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR (CD₃OD) δ 149.8, 142.5, 135.8, 124.9, 119.0, 112.1, 84.3, 82.4,77.1, 71.6, 71.5, 67.4, 65.5, 57.3, 52.7, 46.9, 46.2, 43.7, 42.4, 40.9,30.1, 28.5, 27.0, 26.2, 24.5, 23.2, 17.3, 14.0, 13.1, 7.9

Example 21(S),3(R)-Dihydroxy-20(R)-(5(S),6(S)-Dihydroxy-5-Ethyl-1(S)-Methoxy-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E), 10(19)-Triene

Compound 102

Method: General Procedure 4

Starting material: Compound 402

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.7, 142.9, 133.1, 124.9, 117.2, 111.8, 82.4, 82.0, 75.6,73.8, 71.3, 70.9, 66.8, 56.4, 56.0, 51.5, 45.7, 45.3, 42.9, 40.3, 39.4,29.1, 27.2, 26.5, 26.4, 23.5, 22.1, 17.1, 14.1, 12.6, 7.7

Example 31(S),3(R)-dihydroxy-20(R)-(5(S),6(S)-Dihydroxy-1(S)-Ethoxy-5-Ethyl-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 103

Method: General Procedure 4

Starting material: Compound 403

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.7, 142.9, 133.2, 124.8, 117.2, 111.8, 82.6, 81.9, 75.6,72.3, 71.3, 70.8, 66.8, 64.2, 56.0, 51.5, 45.8, 45.3, 40.2, 39.3, 29.1,27.2, 26.4, 26.3, 23.5, 22.2, 17.1, 15.2, 14.2, 12.7, 7.7

Example 41(S),3(R)-Dihydroxy-20(R)-(5(S),6(S)-Dihydroxy-5-Ethyl-1(S)-(1-n-Propyloxy)-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 104

Method: General Procedure 4

Starting material: Compound 404

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.7, 143.0, 133.0, 125.0, 117.1, 111.8, 82.9, 81.7, 75.6,72.4, 71.3, 70.9, 70.5, 66.9, 56.0, 51.5, 45.8, 45.3, 42.9, 40.3, 39.5,29.1, 27.2, 26.5, 26.3, 23.5, 23.1, 22.2, 17.1, 14.2, 12.7, 10.9, 7.7

Example 51(S),3(R)-Dihydroxy-20(R)-(1(S)-Benzyloxy-5(S),6(S)-Dihydroxy-5-Ethyl-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 105

Method: General Procedure 4

Starting material: Compound 405

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.7, 142.9, 138.5, 133.1, 128.2, 127.7, 127.4, 124.9, 117.1,111.8, 82.6, 82.2, 75.6, 71.9, 71.3, 70.8, 70.4, 66.8, 56.0, 51.5, 45.8,45.3, 42.9, 40.3, 39.4, 29.0, 27.2, 26.4, 26.3, 23.5, 22.1, 20.8, 17.1,12.7, 7.7

Example 61(S),3(R)-Dihydroxy-20(R)-(5(R),6(S)-Dihydroxy-1(S)-Ethoxy-5-Ethyl-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 106

Method: General Procedure 4

Starting material: Compound 407

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.7, 142.8, 133.2, 124.8, 117.2, 111.8, 82.9, 81.9, 75.4,72.3, 71.2, 70.8, 66.8, 64.2, 56.0, 51.5, 45.8, 45.2, 42.9, 40.2, 39.3,29.2, 29.1, 26.2, 25.1, 23.5, 22.2, 17.1, 15.2, 14.2, 12.7, 7.5

Example 71(S),3(R)-Dihydroxy-20(R)-(5(R),6(R)-Dihydroxy-1(S)-Ethoxy-5-Ethyl-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 107

Method: General Procedure 4

Starting material: Compound 409

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ147.7, 142.9, 133.2, 124.9, 117.2, 111.8, 82.6, 81.9, 75.6,72.3, 71.3, 70.8, 66.8, 64.2, 56.0, 51.5, 45.8, 45.2, 42.8, 40.2, 39.3,29.1, 27.2, 26.4, 26.3, 23.5, 22.1, 17.1, 15.2, 14.2, 12.7, 7.7

Example 81(S),3(R)-Dihydroxy-20(R)-(5(S),6(R)-Dihydroxy-1(S)-Ethoxy-5-ethyl-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7E),10(19)-Triene

Compound 108

Method: General Procedure 4

Starting material: Compound 411

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.7, 142.8, 133.2, 124.8, 117.2, 111.8, 83.0, 81.9, 75.4,72.3, 71.2, 70.8, 66.8, 64.2, 56.0, 51.5, 45.8, 45.2, 42.9, 40.2, 39.3,29.3, 29.1, 26.3, 25.1, 23.5, 22.1, 17.1, 15.2, 14.2, 12.7, 7.5

Example 91(S),3(R)-Dihydroxy-20R)-(4-ethyl-1(S),4(S),5(S)-Trihydroxy-2-Hexyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 109

Method: General Procedure 10

Starting material: Compound 412

Chromatography eluant: ethyl acetate.

¹H NMR (CD₃OD) δ 6.32 (d,1H), 6.08 (d,1H), 5.28 (d,1H), 4.90 (d,1H),4.57 (d,1H), 4.35 (t,1H), 4.12 (m,1H), 3.63 (q,1H), 2.86 (dd,1H), 2.55(dd,1H), 2.25 (dd,1H), 2.0-1.0 (m,16H), 1.27 (d,3H), 1.23 (d,3H), 1.05(t,3H), 0.56 (s,3H)

Example 101(S),3(R)-Dihydroxy-20(R)-(4(S),5(S)-Dihydroxy-1(S)-Ethoxy-4-Ethyl-2-Hexyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 111

Method: General Procedure 10

Starting material: Compound 413

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹H NMR δ 6.37 (d,1H), 6.01 (d,1H), 5.32 (t,1H), 5.00 (d,1H), 4.42(t,1H), 4.41 (m,2H), 3.81 (q,1H), 3.73 (m,1H), 3.31 (m,1H), 2.83(dd,1H), 2.59 (dd,1H), 2.31 (m,2H), 2.1-1.0 (m,14H), 1.25 (d,3H), 1.21(t,3H), 1.08 (t,3H), 1.02 (d,3H), 0.55 (s,5H)

Example 111(S),3(R)-Dihydroxy-20(R)-(4-Ethyl-1(S),4(R),5(S)-Trihydroxy-2-Hexyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 114

Method: General Procedure 10

Starting material: Compound 414

Chromatography eluant: ethyl acetate.

¹³C NMR δ (CD₃OD)149.9, 142.4, 135.8, 130.2, 124.9, 119.1, 112.1, 73.5,71.5, 67.4, 65.3, 61.6, 57.3, 52.7, 46.9, 46.2, 43.8, 42.2, 40.9, 32.1,30.1, 27.1, 24.6, 23.2, 18.3, 14.5, 14.0, 13.0, 9.0

Example 121(S),3(R)-Dihydroxy-20(R)-(4(R),5(S)-Dihydroxy-1(S)-Ethoxy-4-Ethyl-2-Hexyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 116

Method: General Procedure 10

Starting material: Compound 415

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.6, 142.9, 133.0, 125.0, 117.1, 111.9, 85.8, 85.3, 75.5,73.2, 72.0, 70.9, 66.9, 64.3, 56.0, 51.5, 45.7, 45.3, 42.9, 40.2, 39.4,31.0, 29.1, 26.4, 23.5, 22.1, 18.5, 15.2, 14.2, 12.7, 8.3

Example 13 Capsules Containing Compound 103

Compound 103 was dissolved in arachis oil to a final concentration of 1μg of Compound 103/ml oil. 10 Parts by weight of gelatine, 5 parts byweight glycerine, 0.08 parts by weight potassium sorbate, and 14 partsby weight distilled water were mixed together with heating and formedinto soft gelatine capsules. These were then filled each with 100 μl ofCompound 103 in oil solution, such that each capsule contained 0.1 μg ofCompound 103.

Example 14 Dermatological Cream Containing Compound 102

In 1 g almond oil was dissolved 0.05 mg of Compound 102. To thissolution was added 40 g of mineral oil and 20 g of self-emulsifyingbeeswax. The mixture was heated to liquefy. After the addition of 40 mlhot water, the mixture was mixed well. The resulting cream containsapproximately 0.5 μg of Compound 102 per gram of cream.

EXAMPLE 15 Injection Fluid Containing Compound 108 Compound 108 (activesubstance) 10 μg Disodium phosphate dihydrate (buffer) 5.4 mg Sodiumdihydrogen phosphate dihydrate (buffer) 2 mg Sodium chloride 0.8 mgSodium ascorbate (antioxidant) 5 mg Solutol ® HS 15 from BASF(solubilizer) 5 mg Water for injection ad 1 ml

Solutol® HS 15 is dissolved in the water for injection by heating it toa temperature of at the most 80° C. A cover of nitrogen is applied. Thebuffer substances and the sodium chloride are added and then thesolution is cooled to at the most 30° C. Then sodium ascorbate is addedand, finally, compound 108 is dissolved in the solution obtained.

The solution is subjected to sterile filtration and is autoclaved at anappropriate time-temperature condition.

Example 161(S),3(R)-Dihydroxy-20(R)-(1(S),5(R/S)-Dihydroxy-5-Ethyl-6(S)-Fluoro-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 157

Method: General Procedure 10

Starting material: Compound 416

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

Example 17 1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy5-ethyl-6(S)-Fluoro-2-Heptyn-5(R/S)-Hydroxy1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 158

Method: General Procedure 10

Starting material: Compound 417

Chromatography eluant: 50% petroleum ether in ethyl acetate.

Example 181(S),3(R)-Dihydroxy-20(R)-(1(S),5(R/S)-Dihydroxy-5-Ethyl-6(R)-Fluoro-2-Heptyn-1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 159

Method: General Procedure 10

Starting material: Compound 418

Chromatography eluant: 50% to 0% petroleum ether in ethyl acetate.

¹³C NMR δ 147.7, 142.7, 133.2, 134.9, 117.3, 111.8, 93.4, 84.2, 88.5,74.5, 78.9, 66.9, 64.8, 56.0, 51.5, 45.7, 45.3, 42.9, 41.0, 39.6, 29.0,28.0, 26.4, 25.3, 23.4, 22.1, 14.9, 13.3, 12.6, 7.5

Example 19 1(S),3(R)-Dihydroxy-20(R)-(1(S)-Ethoxy5-Ethyl-6(R)-Fluoro-2-Heptyn-5(R/S)-Hydroxy1-Yl)-9,10-Secopregna-5(Z),7(E),10(19)-Triene

Compound 160

Method: General Procedure 10

Starting material: Compound 419

Chromatography eluant: 50% petroleum ether in ethyl acetate.

¹H NMR δ 6.37 (d,1H), 6.02 (d,1H), 5.33 (s,1H), 5.00 (s,1H), 4.68(dq,1H), 4.43 (m,1H), 4.23 (m,1H), 4.13 (s,1H), 3.73 (m,1H), 3.30(m,1H), 2.84 (dd,1H), 2.58 (m,2H), 2.40 (dd,1H), 2.31 (dd,1H), 2.05-1.20(m,19H), 1.38 (dd,3H), 1.20 (t,3H),1.01 (d,3H), 0.96 (t,3H), 0.54 (s,3H)

What is claimed is:
 1. A compound of the formula I

wherein R represents hydrogen, or R represents (C₁-C₆)alkyl, phenyl, or(C₇-C₉)aralkyl, optionally substituted with one or more groups selectedfrom (C₁-C₃)alkyl, F, and phenyl; n is an integer having the value 0, 1,or 2; and X represents hydroxy or halogen.
 2. A compound according toclaim 1 wherein R is selected from the group consisting of methyl,ethyl, propyl, isopropyl, benzyl, ortho methylbenzyl, meta methylbenzyl,and para methylbenzyl.
 3. A compound according to claim 1 wherein n is 0or
 1. 4. A compound according to claim 1 wherein X represents asubstituent selected from the group consisting of OH, F, and Cl.
 5. Acompound according to claim 1 having the configuration 22(S), 25(S),26(S) or 22(S), 25(S), 26(R).
 6. A compound according to claim 1selected from the group consisting of1(S),3(R)-Dihydroxy-20(R)-(5-ethyl-1(S),5(S),6(S)-trihydroxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 101),1(S),3(R)-Dihydroxy-20(R)-(5(S),6(S)-dihydroxy-5-ethyl-1(S)-methoxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 102),1(S),3(R)-Dihydroxy-20(R)-5(S),6(S)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 103),1(S),3(R)-Dihydroxy-20(R)-(5(S),6(S)-dihydroxy-5-ethyl-1(S)-(1-propyloxy)-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 104),1(S),3(R)-Dihydroxy-20(R)-(1(S)-benzylyloxy-5(S),6(S)-dihydroxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 105),1(S),3(R)-Dihydroxy-20(R)-(5(R),6(S)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 106),1(S),3(R)-Dihydroxy-20(R)-(5(R),6(R)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 107),1(S),3(R)-Dihydroxy-20(R)-(5(R),6(R)-dihydroxy-1(S)-ethoxy-5-ethyl-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 108),(S),3(R)-Dihydroxy-20(R)-(4-ethyl-1(S),4(S),5(S)-trihydroxy-2-hexyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound (109),(S),3(R)-Dihydroxy-20(R)-(4(S),5(S)-dihydroxy-1(S)-ethoxy-4-ethyl-2-hexyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 111),1(S),3(R)-Dihydroxy-20(R)-(4-ethyl-1(S),4(R),5(S)-trihydroxy-2-hexyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 114),1(S),3(R)-Dihydroxy-20(R)-(4(R),5(S)-dihydroxy-1(S)-ethoxy-4-ethyl-2-hexyn-1-yl)9,10-secopregna-5(Z),7(E),10(19)-triene (Compound 116),1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy-5-ethyl-6(S)-fluoro-5(S)-hydroxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 149), and1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy-5-ethyl-6(R)-fluoro-5(S)-hydroxy-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 150),1(S),3(R)-Dihydroxy-20(R)-(1(S),5(R/S)-dihydroxy-5-ethyl-6(S)-fluoro-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 157) 1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy5-ethyl-6(S)-fluoro-2-heptyn-5(R/S)-hydroxy1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 158)1(S),3(R)-Dihydroxy-20(R)-(1(S),5(R/S)-dihydroxy-5-ethyl-6(R)-fluoro-2-heptyn-1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 159) 1(S),3(R)-Dihydroxy-20(R)-(1(S)-ethoxy5-ethyl-6(R)-fluoro-2-peptyn-5(R/S)-hydroxy1-yl)-9,10-secopregna-5(Z),7(E),10(19)-triene (compound 160).
 7. Amethod for producing a compound of formula I of claim 1 characterized ina) reacting1(S),3(R)-bis-(tert-butyldimethylsilyloxy)-20(R)-formyl-9,10-secopregna-5(E),7(E),10(19)-triene, with an organometallic reagent derived from the sidechain building block HC≡C—(CH₂)_(n)—C(C₂H₅)(O—PG¹)(CHX¹—CH₃), wherein nand R have the meanings specified above; X¹ represents O—PG², fluorine,chlorine, bromine, or iodine; PG¹ and PG² both represent the same ordifferent group selected from trimethylsilyl andtert-butyl-dimethylsilyl; or PG¹ and PG² together form one bifunctionalisopropylidene ketal protective group; to form a compound of formulaIII, and b) optionally separating from diastereoisomers the compoundformed in step a), c) subjecting the compound formed in step a) or stepb) to triplet-sensitized photo-isomerization to the 5(Z) isomer, d)optionally alkylating the compound formed in step c) at the 22-hydroxygroup with a (C₁-C₆)alkyl or (C₇-C₉)aralkyl bromide or iodide in thepresence of a base and a phase transfer catalyst, and e) deprotectingthe compound formed in step c) or step d) to form a compound of formulaI.
 8. A pharmaceutical composition containing an effective amount of oneor more of the compounds of claim 1, together with pharmaceuticallyacceptable, non-toxic carriers and/or auxiliary agents.
 9. A compoundaccording to claim 1 wherein n is
 1. 10. A method of inhibitingproliferation of leukaemia cells, breast cancer cells and skin cells,the method comprising contacting said cells with an effective amount ofa compound according to claim 1, optionally in combination with othertherapeutically active compounds.
 11. A compound according to claim 1wherein the compound is a diastereoisomer of formula I.
 12. A compoundaccording to claim 1 wherein the compound is a diastereoisomer offormula I in pure form.
 13. A compound according to claim 1 wherein thecompound comprises a mixture of diastereoisomers of formula I.
 14. Acompound according to claim 3 wherein n is 1.